Targeted Integration Through Transformation of Hydroxyurea-Arrested Cells
1Ginkgo Bioworks, Boston, MA, USA. vtsakraklides@ginkgobioworks.com.
Methods in Molecular Biology (Clifton, N.J.)
|April 13, 2021
Abstract:
Homologous recombination is required to specifically target DNA to a desired genomic locus. Non-homologous end joining is the predominant form of recombination in Yarrowia lipolytica. Transformation of this organism with linear DNA therefore results in random integration of the introduced DNA into the genome. In this protocol, hydroxyurea-mediated cell cycle arrest is applied to significantly increase the rate of homologous recombination during transformation and enhance targeted integration.
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